AC10 human ventricular cardiomyocytes (PTA-1501) and DMEM:F12 medium (supplemented with 2.5 mM glutamine, 15 mM HEPES, 0.5mM sodium pyruvate, and 1200 mg/L sodium bicarbonate) were obtained from the ATCC (Manassas, VA). Characterized fetal bovine serum (FBS) and Dulbecco’s phosphate-buffered saline without Ca2+ and Mg2+ (PBS), and thioflavin T (ThT) were purchased from Thermo Scientific (Waltham, MA). Hank’s balanced salt solution, 100x penicillin-streptomycin solution, L-glutamine, trypsin, and EDTA were purchased from Corning Life Sciences (Tewksbury, MA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dichloro-dihydro-fluorescein diacetate (DCFH-DA), elastin, crystal violet, and anhydrous dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO). Cellular ATP quantitation was performed with the CellTiter-Glo Luminescent Cell Viability Assay kit from Promega (Madison, WI) according to the manufacturer’s directions. Hoescht 33342, Alexa Fluor 488 NHS Ester (succinimidyl ester), and Alexa Fluor 647 phalloidin were purchased from Life Technologies (Grand Island, NY). BD Cytofix was from Becton-Dickinson (Franklin Lakes, NJ). 5-chloromethylfluorescein diacetate (CMFDA) was purchased from Genecopoeia (Rockville, MD).
Dmem f12 medium
Manufactured by American Type Culture Collection
Sourced in United States
DMEM:F12 medium is a cell culture medium that supports the growth and maintenance of a variety of cell types. It is a combination of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture. The medium provides a balanced formulation of nutrients, vitamins, and salts to support cell proliferation and survival in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (6 mentions)
Fetal bovine serum (fbs), by Merck Group (5 mentions)
Hct116, by American Type Culture Collection (5 mentions)
Arpe 19, by American Type Culture Collection (5 mentions)
Hydrocortisone, by Merck Group (4 mentions)
Rpmi 1640 medium, by American Type Culture Collection (4 mentions)
Sw620, by American Type Culture Collection (4 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Insulin, by Merck Group (3 mentions)
Fetal calf serum (fcs), by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Its premix, by Corning (3 mentions)
Nu serum, by Corning (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Pen strep, by Thermo Fisher Scientific (3 mentions)
61 protocols using dmem f12 medium
1
Characterization of Human Ventricular Cardiomyocytes
2
Isolation and Culture of Skin Fibroblasts
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Fibroblasts were obtained by digesting the skin punch biopsy samples in dispase and collagenase IV (both from STEMCELL Technologies Inc., Cambridge, MA, USA) enzymes mixture (2.5 hours at 37°C) and undigested tissues were filtered through a 100 μm cell strainer. The cell suspensions were diluted in DMEM/F12 medium (ATCC) and centrifuged at 300 g for 8 min. Collected cells were grown in DMEM/F12 medium supplemented with 10% foetal bovine serum (FBS) (Sigma-Aldrich, Saint-Louis, MO, USA), penicillin/streptomycin and amphotericin B (Sigma-Aldrich) at 37°C in a humidified 5% CO2 incubator.
3
Culturing and Collecting Conditioned Media
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On day 1, hUTCs or ARPE-19 cells were thawed and plated in a 6-well plate at 0.288 × 106 cells/2.4 mL/well in DMEM-LG medium containing 15% (v/v) FBS (HyClone, Logan, UT) and 4 mM L-glutamine (Gibco, Grand Island, NY). On day 2, medium was aspirated and replenished with DMEM/F12 medium (ATCC) containing 10% v/v FBS, 50 U/mL penicillin (Invitrogen), and 50 μg/mL streptomycin (Invitrogen). Cells remained in these culture conditions for 48 h, after which CM from hUTCs or ARPE-19 cells was collected to be used for experiments or frozen at −70°C for future use.
4
Cell Culture and Viability Assay Protocol
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The cell lines used in the present study were: HGF—human primary gingival fibroblasts (ATCC® PCS-201-018™), SCC-4—human squamous cell carcinoma cell line (ATCC® CRL-1624™), HaCaT—immortalized human keratinocytes (CLS Cell Lines Service GmbH), and A375—human melanoma cells (ATCC® CRL-1619™).
For cell culture and cell viability assay, the following reagents were needed: specific culture medium—Fibroblast Basal Medium (ATCC PCS-201-030) and Fibroblast growth kit—low serum (ATCC PCS-201-041) for HGF cells, DMEM:F12 Medium (ATCC® 30-2006™)—for SCC-4 cells were acquired from ATCC, and Dulbecco’s Modified Eagle Medium (DMEM) high glucose −4.5 g/L for HaCaT and A375 cells, together with the other reagents used, as: trypsin—EDTA solution, PBS (phosphate saline buffer), fetal bovine serum (FCS), Trypan blue, Alamar blue (resazurin sodium salt) and hydrocortisone were purchased from Sigma Aldrich (Darmstadt, Germany) and Thermo Fisher Scientific (Waltham, MA, USA).
For cell culture and cell viability assay, the following reagents were needed: specific culture medium—Fibroblast Basal Medium (ATCC PCS-201-030) and Fibroblast growth kit—low serum (ATCC PCS-201-041) for HGF cells, DMEM:F12 Medium (ATCC® 30-2006™)—for SCC-4 cells were acquired from ATCC, and Dulbecco’s Modified Eagle Medium (DMEM) high glucose −4.5 g/L for HaCaT and A375 cells, together with the other reagents used, as: trypsin—EDTA solution, PBS (phosphate saline buffer), fetal bovine serum (FCS), Trypan blue, Alamar blue (resazurin sodium salt) and hydrocortisone were purchased from Sigma Aldrich (Darmstadt, Germany) and Thermo Fisher Scientific (Waltham, MA, USA).
5
Culturing Breast Cancer Cell Lines
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The human breast adenocarcinoma cell lines MCF-7 (ATCC® HTB-22) and MDA-MB-231 (ATCC® HTB-26) and the breast epithelial cells MCF-10A (ATCC® CRL-10317) were acquired from the American Type Culture Collection (ATCC). MCF7 cells were cultured in Eagle's Minimum Essential Medium (EMEM; ATCC) and MDA-MB-231 cells were cultured in high glucose Dulbecco's Modified Eagle's Medium (DMEM; Sigma-Aldrich), whereas MCF-10A cells were cultured in 1 : 1 mixture DMEM: F-12 medium (ATCC) supplemented with 20 ng/mL epidermal growth factor (EGF; Gibco, Thermo Fisher Scientific), 0.01 ng/mL insulin (Sigma-Aldrich), 500 ng/mL hydrocortisone (Sigma-Aldrich), and 5% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific). Tumorigenic cell lines were supplemented with 10% FCS. 1% penicillin/streptomycin mixture (Pen/Strep, 10,000 IU/ml; Sigma-Aldrich) was added in each cell culture medium to avoid a possible fungal/microbial contamination. Standard conditions were used for cell culture—37°C and humidified atmosphere containing 5% CO2, as previously described before [34 (link), 35 (link)].
6
Mammalian Cell Culture and Transfection
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HeLa cell was cultured in Dulbecco’s modified Eagle’s medium (Sigma Chemical Co.) supplemented with 10% heat-inactivated calf serum at 37 °C under a 5% CO2 atmosphere. Transfection in HeLa cells with pCI-neo expression vectors was performed by using PEI MAX (Polysciences, Inc, PA, USA) or HilyMax (Dojindo, Kyoto, Japan) according to the protocols supplied by the manufacturers. At 24 h after plasmid transfection, the cells were harvested and subjected to immunological analysis, unless otherwise noted.
Human telomerase reverse transcriptase retinal pigment epithelium 1 (hTERT-RPE1) cells were purchased from American Type Culture Collection (ATCC), and were cultured according to the manual provided. Briefly, cells were cultured at 37°C under 5% CO2 in Dulbecco’s modified Eagle medium–nutrient mixture F-12 (DMEM/F12) medium (ATCC) supplemented with 10% fetal bovine serum (FBS).
Human telomerase reverse transcriptase retinal pigment epithelium 1 (hTERT-RPE1) cells were purchased from American Type Culture Collection (ATCC), and were cultured according to the manual provided. Briefly, cells were cultured at 37°C under 5% CO2 in Dulbecco’s modified Eagle medium–nutrient mixture F-12 (DMEM/F12) medium (ATCC) supplemented with 10% fetal bovine serum (FBS).
7
Neuroblastoma Cell Line Arachidonic Acid Uptake
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Human-derived neuroblastoma SH-SY5Y cells (ATCC, Manassas, VA) were given DMEM-F12 medium (ATCC) (with 20% fetal bovine serum (FBS) and penicillin-streptomycin added) and cultured in a temperature-controlled CO2 incubator at 37°C. In assays treating SH-SY5Y cells with LPS or TNFα, overnight serum starvation was followed by exposure either with LPS (20 μg/ml) or TNFα (20 ng/ml) or LPS plus celastrol (100 nM) in DMEM-F12 (with 0.5% FBS and no antibiotics added). SH-SY5Y cells were subjected to pretreatment with celastrol (100 nM) for 5 h before LPS treatment. After 48 h of LPS or TNFα or LPS plus celastrol treatment, 14C-AA uptake was performed in vitro [33 (link), 34 (link)]. Briefly, SH-SY5Y cells were incubated (30 min) with 14C-AA (0.1 μCi) in Krebs-Ringer (KR) buffer at 37°C in a water bath, and then, sample lysates were prepared for radioactivity determination using a liquid scintillation counter [33 (link), 34 (link)].
8
Estrogen and Bisphenol A Effects on Prostate Cancer
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The LNCaP and PC3 cell lines were a kind gift from Dr. Shuk-Mei Ho (University of Cincinnati, Cincinnati, OH). The LNCaP cell line was maintained in RPMI/1640 medium (Life Technologies) supplemented with 2 mM/L l -glutamine and 1 mM sodium pyruvate (Gibco). PC3 cells were maintained in DMEM/F12 medium (ATCC). Both medium were supplemented with 10% heat inactivated fetal bovine serum (Atlanta Biologicals) and 100 units/mL of penicillin–streptomycin (Life Technologies). Cells were cultured at 37 °C in a 5% CO2 heated incubator.
17β-Estradiol, bisphenol A, and ICI 182,780 (Fulvestrant) were purchased from Sigma–Aldrich. Cells were treated with 1, 5, or 10 nM estradiol or 10, 25, or 50 nM bisphenol A for 24 h. Where indicated, the ER inhibitor ICI was added at 10 μM at the same time as the estradiol or BPA. All treatments were for 24 h and 95% ethanol (0.1%) was used as the vehicle control.
17β-Estradiol, bisphenol A, and ICI 182,780 (Fulvestrant) were purchased from Sigma–Aldrich. Cells were treated with 1, 5, or 10 nM estradiol or 10, 25, or 50 nM bisphenol A for 24 h. Where indicated, the ER inhibitor ICI was added at 10 μM at the same time as the estradiol or BPA. All treatments were for 24 h and 95% ethanol (0.1%) was used as the vehicle control.
9
Culturing of Human Colon Cell Lines
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Normal human colon epithelial cell line (FHC) and five human CRC cell lines (HCT-116, HT29, LoVo, SW480, and SW620) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). FHC cells were cultured in DMEM/F12 medium (ATCC) containing 25 mM HEPES, 10 ng/ml cholera toxin, 5 μg/ml insulin and transferrin, and 100 ng/ml hydrocortisone. LoVo cells, HCT-116 and HT29 cells, and SW480 and SW620 cells were grown in F-12K medium, McCoy’s 5a medium modified, and Leibovitz’s L-15 medium (all ATCC), respectively. All of the media were supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA). Cells were maintained at 37°C in a humidified incubator under 5% CO2 condition10 (link).
10
SARS-CoV-2 Neutralization Assay in Vero E6 and Calu-3 Cells
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Infection and APN01 mediated viral neutralisation assays were conducted at the Integrated Research Facility at Fort Detrick (IRF-Frederick) of the National Institute of Allergy and Infectious Diseases (NIAID) or at the Department of Laboratory Medicine (Unit of Clinical Microbiology) of the Karolinska Institutet and Karolinska University Hospital. Vero E6 cells were cultured in DMEM medium (Gibco, Gaithersburg, MD, USA or Thermofisher) containing 10% fetal bovine serum (FBS). Calu-3 cells (HTB-55; American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in DMEM F12 Medium (ATCC) with 20% FBS.
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